基于DNA条形码技术的粉螨种类鉴定研究
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摘要
目的建立常见粉螨DNA条形码鉴定方法。方法对2017年3-12月采自安徽省部分地区的面粉、中药材、饲料等储藏物中的粉螨进行形态学鉴定,获得相应种类线粒体细胞色素C氧化酶亚基Ⅰ(COⅠ)基因序列作为标准片段。同时,通过美国国立生物技术信息中心检索已知粉螨COⅠ基因序列,应用DNAStar和MEGA 6.0软件进行序列分析,使用Neighbor-Joining(NJ)法构建系统发育树。结果经形态学鉴定出7种粉螨,分别为粗脚粉螨、椭圆食粉螨、腐食酪螨、害嗜鳞螨、拱殖嗜渣螨、棕脊足螨和粉尘螨。DNA条形码技术鉴定结果与形态学鉴定结果一致,种间遗传距离为0.058～0.323,种内遗传距离为0.000～0.015。NJ系统进化树中,粉螨聚类分析结果与形态学鉴定的物种分类地位一致。结论 DNA条形码可作为形态学鉴定方法的补充,对粉螨种类进行准确快速鉴定。
Objective To establish a DNA barcoding method for identification of common species of acaroid mites.Methods From March to December, 2017, acaroid mites were collected from flour, Chinese medicinal materials, and normal diet in different parts of Anhui province, China, for morphological identification. Gene sequences of mitochondrial cytochrome C oxidase subunitⅠ(COⅠ) from corresponding species were used as standard fragments. The COⅠ gene sequences from known acaroid mites were collected from the National Center for Biotechnology Information and analyzed using DNAStar and MEGA 6.0. A phylogenetic tree was constructed using the neighbor-joining(NJ) algorithm. Results Morphological identification revealed 7 species of acaroid mites: Acarus siro, Aleuroglyphus ovatus, Tyrophagus putrescentiae,Lepidoglyphus destructor, Gohieria fusca, Chortoglyphus arcuatus, and Dermatophagoides farinae. The results of DNA barcoding identification were consistent with the morphological results. The interspecific genetic distance was 0.058-0.323,while the intraspecific genetic distance was 0.000-0.015. In the NJ phylogenetic tree, the cluster analysis results of acaroid mites were consistent with the taxonomic status by morphological identification. Conclusion DNA barcoding can be used as a supplement to morphological identification, which provides fast and accurate identification of the species of acaroid mites.
Objective To establish a DNA barcoding method for identification of common species of acaroid mites.Methods From March to December, 2017, acaroid mites were collected from flour, Chinese medicinal materials, and normal diet in different parts of Anhui province, China, for morphological identification. Gene sequences of mitochondrial cytochrome C oxidase subunitⅠ(COⅠ) from corresponding species were used as standard fragments. The COⅠ gene sequences from known acaroid mites were collected from the National Center for Biotechnology Information and analyzed using DNAStar and MEGA 6.0. A phylogenetic tree was constructed using the neighbor-joining(NJ) algorithm. Results Morphological identification revealed 7 species of acaroid mites: Acarus siro, Aleuroglyphus ovatus, Tyrophagus putrescentiae,Lepidoglyphus destructor, Gohieria fusca, Chortoglyphus arcuatus, and Dermatophagoides farinae. The results of DNA barcoding identification were consistent with the morphological results. The interspecific genetic distance was 0.058-0.323,while the intraspecific genetic distance was 0.000-0.015. In the NJ phylogenetic tree, the cluster analysis results of acaroid mites were consistent with the taxonomic status by morphological identification. Conclusion DNA barcoding can be used as a supplement to morphological identification, which provides fast and accurate identification of the species of acaroid mites.
引文
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[11]Kumar S,Nei M,Dudley J,et al.MEGA:a biologist-centric software for evolutionary analysis of DNA and protein sequences[J].Brief Bioinform,2008,9(4):299-306.DOI:10.1093/bib/bbn017.
[12]Yang BH,Li CP.The full-length mitochondrial r DNA sequence of Tyrophagus longior(Astigmata:Acaridae)and phylogeny of astigmatid mites based on mitochondrial r DNA sequence[J].JStored Prod Res,2017,71:5-13.DOI:10.1016/j.jspr.2016.12.004.
[13]Hebert PDN,Stoeckle MY,Zemlak TS,et al.Identification of birds through DNA barcodes[J].PLo S Biol,2004,2(10):e312.DOI:10.1371/journal.pbio.0020312.
[14]Beroiz B,Couso-Ferrer F,Ortego F,et al.Mite species identification in the production of allergenic extracts for clinical use and in environmental samples by ribosomal DNA amplification[J].Med Vet Entomol,2014,28(3):287-296.DOI:10.1111/mve.12052.2018-11-19
[2]Stala?s A,Moro?ko-Bi?evska I.Species identification,host range and diversity of Cecidophyopsis mites(Acari:Trombidiformes)infesting Ribes in Latvia[J].Exp Appl Acarol,2016,69(2):129-153.DOI:10.1007/s10493-016-0024-7.
[3]Hubert J,Kopecky J,Sagova-Mareckova M,et al.Assessment of bacterial communities in thirteen species of laboratory-cultured domestic mites(Acari:Acaridida)[J].J Econ Entomol,2016,109(4):1887-1896.DOI:10.1093/jee/tow089.
[4]Vargas M,García-Varela M,Laclette JP,et al.Application of ITS-2 sequences as markers for identification and phylogenetic inference within the genus Geomylichus(Acari:Listrophoridae)[J].Exp Appl Acarol,2005,35(3):223-238.DOI:10.1007/s10493-004-2761-2.
[5]Zuo YJ,Chen ZJ,Kondo K,et al.DNA barcoding of Panax species[J].Planta Med,2011,77(2):182-187.DOI:10.1055/s-0030-1250166.
[6]Hebert PDN,Cywinska A,Ball SL,et al.Biological identifications through DNA barcodes[J].Proc Biol Sci,2003,270(1512):313-321.DOI:10.1098/rspb.2002.2218.
[7]Cruickshank RH.Molecular markers for the phylogenetics of mites and ticks[J].Syst Appl Acarol,2002,7:3-14.DOI:10.11158/saa.7.1.1.
[8]Yang B,Cai JL,Cheng XJ.Identification of astigmatid mites using ITS2 and COⅠregions[J].Parasitol Res,2011,108(2):497-503.DOI:10.1007/s00436-010-2153-y.
[9]Jia FX,Yang MS,Yang WJ,et al.Influence of continuous high temperature conditions on Wolbachia infection frequency and the fitness of Liposcelis tricolor(Psocoptera:Liposcelididae)[J].Environ Entomol,2009,38(5):1365-1372.DOI:10.1603/022.038.0503.
[10]Webster LMI,Thomas RH,Mc Cormack GP.Molecular systematics of Acarus siro s.lat.,a complex of stored food pests[J].Mol Phylogenet Evol,2004,32(3):817-822.DOI:10.1016/j.ympev.2004.04.005.
[11]Kumar S,Nei M,Dudley J,et al.MEGA:a biologist-centric software for evolutionary analysis of DNA and protein sequences[J].Brief Bioinform,2008,9(4):299-306.DOI:10.1093/bib/bbn017.
[12]Yang BH,Li CP.The full-length mitochondrial r DNA sequence of Tyrophagus longior(Astigmata:Acaridae)and phylogeny of astigmatid mites based on mitochondrial r DNA sequence[J].JStored Prod Res,2017,71:5-13.DOI:10.1016/j.jspr.2016.12.004.
[13]Hebert PDN,Stoeckle MY,Zemlak TS,et al.Identification of birds through DNA barcodes[J].PLo S Biol,2004,2(10):e312.DOI:10.1371/journal.pbio.0020312.
[14]Beroiz B,Couso-Ferrer F,Ortego F,et al.Mite species identification in the production of allergenic extracts for clinical use and in environmental samples by ribosomal DNA amplification[J].Med Vet Entomol,2014,28(3):287-296.DOI:10.1111/mve.12052.2018-11-19